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EnzyChrom™ Cholesterol Assay Kit


  • For quantitative determination of cholesterol and evaluation of drug effects on cholesterol metabolism.

Key Features

  • Sensitive and accurate. The detection limit of 5 mg/dL, linearity up to 300 mg/dL cholesterol in a 96-well plate assay.
  • Convenient. Room temperature assay. No 37°C heater is needed.
  • High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.


  • OD340nm


  • Serum, plasma etc


  • All


  • 40 min


  • 100 tests

Detection Limit

  • 5 mg/dL

Shelf Life

  • 6 months

More Details

CHOLESTEROL is a sterol and lipid present in the cell membranes, and is transported in the bloodstream of all animals. It is used to form cell membranes and hormones and plays important roles in cell signaling processes. Elevated levels (hypercholesterolemia) have been associated with cardiovascular diseases such as atherosclerosis; whereas, low levels (hypocholesterolemia) may be linked to depression, cancer and cerebral haemorrhage. Simple, direct and automation-ready procedures for measuring cholesterol are very desirable. BioAssay Systems EnzyChrom™ Cholesterol Assay is based on cholesterol esterase hydrolysis of cholesterol esters to form free cholesterol and cholesterol dehydrogenase catalyzed conversion of cholesterol to cholest-4-ene-3-one, in which NAD is reduced to NADH. The optical density of the formed NADH at 340 nm is directly proportionate to the cholesterol concentration in the sample.

I would like to know whether egg yolk can be used for such a kit. If this doesn’t work, can you suggest us another protocol, please?

Egg yolk cholesterol extraction protocol:

1. Prepare extraction buffer with 5 vol isopropanol, 2 vol water, 2 vol Triton X-100.

In a 1.5mL Eppendorf tube, mix 50µL yolk with 450 µL of the extraction buffer. Vortex for at least 30 sec.

3. Centrifuge 4 min at 14,000rpm on a table centrifuge.

4. Remove supernatant and perform cholesterol assay.

What is the principle of the ECCH-100 cholesterol assay?

Cholesterol esterase cleaves esters into cholesterol, which is oxidized to form NADH in the presence of cholesterol dehydrogenase. The produced NADH is measured colorimetrically.

Will the assay detect the cholesterol in the lipoproteins, or only free-cholesterol in the serum?

The assay will detect total serum cholesterol, i.e. protein-bound serum (VLDL/LDL/HDL) and free cholesterol.<

Jiang, S.,et al. (2021). Associations of Circulating Irisin with FNDC5 Expression in Fat and Muscle in Type 1 and Type 2 Diabetic Mice. Biomolecules. 11(2): 322. Assay: Cholesterol in mouse plasma.

Moorhead, S. G., et al. (2020). Variation of body condition and plasma energy substrates with life stage, sex, and season in wild-sampled nurse sharks Ginglymostoma cirratum . Journal of Fish Biology. jfb: 14612. Assay: Cholesterol in shark plasma.

Torres, E., et al. (2020). Serum metabolites and body condition score associated with metritis, endometritis, ketosis, and mastitis in Holstein cows. Pesquisa Agropecuaria Brasileira. 55: e01308. Assay: Cholesterol in cow serum.

Jeffrey, J. D., et al. (2019). Physiological status of silver carp (Hypophthalmichthys molitrix) in the Illinois River: An assessment of fish at the leading edge of the invasion front. Comparative Biochemistry and Physiology. Part D, Genomics & Proteomics. 32, 100614. Assay: Cholesterol in silver carp plasma.

Shin, M. K., et al. (2020). Capsaicin suppresses liver fat accumulation in high-fat diet-induced NAFLD mice. Animal Cells and Systems. 24(4): 214-219. Assay: Cholesterol in mouse plasma.

Basiel, B. L., et al. (2020). Cholesterol deficiency carriers have lowered serum cholesterol and perform well at an elite cattle show. JDS Communications. 1(1): 6-9. Assay: Cholesterol in cattle serum.

Yoon, J. A., et al. (2020). Evaluation of changes in body composition and proinflammatory marker levels in ovariectomized rats with induced hyperthyroidism. All Life. 13(1): 99-107. Assay: Cholesterol in rat serum.

Berkes, E., et al. (2020). Potentiated in vitro probiotic activities of lactobacillus fermentum lfqi6 biofilm biomass versus planktonic culture. Probiotics and Antimicrobial Proteins. 12(3): 1097-1114. Assay: Cholesterol in Lactobacillus fermentum LfQi6 culture.

Al-Ziadi, R. E (2017). The effect of Mutagens agent Ethidium bromide on some therapeutic properties for local Iraqi isolate Saccharomyces boulardii. Scientific Journal of Medical Research 1.1: 13-18. Assay: Cholesterol in Saccharomyces boulardii (Yeast) culture medium.

Brooks, Sylwia W., Ava C. Dykes, and Bernard G. Schreurs (2017). A high-cholesterol diet increases 27-hydroxycholesterol and modifies estrogen receptor expression and neurodegeneration in rabbit hippocampus. Journal of Alzheimer’s Disease 56.1 (2017): 185-196. Assay: Cholesterol in rabbits serum.

Hidanah, Sri, et al (2017). Effects of mangosteen peel (Garcinia mangostana) and ginger rhizome (Curcuma xanthorrhiza) on the performance and cholesterol levels of heat-stressed broiler chickens. Pakistan Journal of Nutrition 16.1: 28-32. Assay: Cholesterol in broiler chicks tissues.

Kwan, Billy KY, et al (2017). Marine microalgae as dietary supplements in the culture of juvenile Chinese horseshoe crabs, Tachypleus tridentatus (Xiphosura). Aquaculture research 48.7: 3910-3924. Assay: Cholesterol in horseshoe crabs plasma.

Mellado, Miguel, et al (2017). Effect of quality of colostrum on health, growth and immunoglobulin G concentration in Holstein calves in a hot environment. Animal Science Journal 88.9: 1327-1336. Assay: Cholesterol in calves serum.

Meza-Herrera, Cesar A., et al (2017). Beta-carotene supplementation positively affects selected blood metabolites across time around the onset of puberty in goats. Czech Journal of Animal Science 62.1 (2017): 22-31. Assay: Cholesterol in goats serum.

Ryan, Paul M., et al (2017). Microbiome and metabolome modifying effects of several cardiovascular disease interventions in apo-E-/- mice. Microbiome 5.1: 30. Assay: Cholesterol in mice serum.

Ryan, Paul M., et al (2017). Recombinant incretin-secreting microbe improves metabolic dysfunction in high-fat diet fed rodents. Scientific reports 7.1: 13523. Assay: Cholesterol in rodent serum.

Stanton, Catherine, et al (2017). Exopolysaccharide-producing bacteria, and uses thereof for protecting heart health. U.S. Patent No. 9,808,493. Assay: Cholesterol in mice serum.

Kaliber, M., N. Koluman, and N. Silanikove (2016). Physiological and behavioral basis for the successful adaptation of goats to severe water restriction under hot environmental conditions. animal 10.1: 82-88. Assay: Cholesterol in goats plasma.

King, Gregory D., et al (2016). Watershed-scale land use activities influence the physiological condition of stream fish. Physiological and Biochemical Zoology 89.1: 10-25. Assay: Cholesterol in fish plasma.

Liss, Stephanie A., et al (2016). Physiological consequences of hybridization: early generation backcrossing decreases performance in invasive bigheaded carps. Journal of Freshwater Ecology 31.4: 543-554. Assay: Cholesterol in fish plasma.

Liss, S. et al (2013). Spatial and temporal influences on the physiological condition of invasive silver carp. Conserv Physiol. 1 (1): cot017 Assay: Cholesterol in fish (silver carp) plasma.

Safwat Y, et al (2013). Modulation of skeletal muscle performance and SERCA by exercise and adiponectin gene therapy in insulin-resistant rat. DNA Cell Biol. 32(7):378-85. Assay: Cholesterol in rat plasma.

Kang, NH et al (2012). Modulation of lipid metabolism by mixtures of protamine and chitooligosaccharide through pancreatic lipase inhibitory activity in a rat model. Lab Anim Res ;8(1):31-8. Assay: Cholesterol in rat feces.

Kang, NH et al (2012). Risk of cardiovascular disease is suppressed by dietary supplementation with protamine and chitooligosaccharide in Sprague-Dawley rats. Mol Med Rep. ;7(1):127-33. Assay: Cholesterol in rat feces.

Mellado, M et al (2012). Effect of dietary energy intake and somatotropin administration after weaning on growth rate and semen characteristics of Granadina goat bucks. Turk. J. Vet. Anim. Sci. 36(4): 338-345. Assay: Cholesterol in goat buck sersum.

Schlotter F, Matsumoto Y, Mangner N, Schuler G, Linke A, et al. (2012). Regular Exercise or Changing Diet Does Not Influence Aortic Valve Disease Progression in LDLR Deficient Mice. PLoS ONE 7(5): e37298. Assay: Cholesterol in mice serum.

Se, EY et al (2012). alpha-Lipoic acid reduced weight gain and improved the lipid profile in rats fed with high fat diet. Nutr Res Pract. 6(3):195-200. Assay: Cholesterol in rat plasma.

Henein, HY et al (2011). Effect of adrenomedullin gene delivery on insulin resistance in type 2 diabetic rats. J Adv Res 2(1): 57-64. Assay: Cholesterol in rat serum.

Hoda, Y. et al ( 2010). Effect of adrenomedullin gene delivery on insulin resistance in type 2 diabetic rats. J. Adv. Res. 2:57-64. Assay: Cholesterol in fish serum.

Hoda, Y. et al ( 2010). Effect of adrenomedullin gene delivery on insulin resistance in type 2 diabetic rats. J. Adv. Res. 2:57-64. Assay: Cholesterol in rat serum.

Hu, M et al. (2010) Effect of prolonged starvation on body weight and blood-chemistry in two horseshoe crab species: Tachypleus tridentatus and Carcinoscorpius rotundicauda (Chelicerata: Xiphosura). J. Exp Marine Biol Ecology 395(1-2):112-119. Assay: Cholesterol in crab plasma.

Hu, M et al. (2010). Effect of prolonged starvation on body weight and blood-chemistry in two horseshoe crab species: Tachypleus tridentatus and Carcinoscorpius rotundicauda (Chelicerata: Xiphosura). J Exp Marine Biol Ecol 395(1-2): 112-119. Assay: Cholesterol in Horseshoe crab plasma.

Rubinstein J, et al (2009). Hypercholesterolemia and myocardial function evaluated via tissue doppler imaging. Cardiovasc Ultrasound. 7:56. Assay: Cholesterol in rabbit serum.

To find more recent publications, please click here.

If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.

Cholesterol Assay Kit
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For bulk quote or custom reagents, please email or call 1-510-782-9988 x 1.

Orders are shipped the same day if placed by 2pm PST
Shipping: On Ice
Carrier: Fedex
Delivery: 1-2 days (US), 3-6 days (Intl)
Storage: -20°C upon receipt

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